title: Refolding of the recombinant protein Sm29, a step toward the production of the vaccine candidate against schistosomiasis.
abstract: Schistosomiasis is an important parasitic disease, with about 240 million people infected worldwide. Humans and animals can be infected, imposing an enormous social and economic burden. The only drug available for chemotherapy, praziquantel, does not control reinfections, and an efficient vaccine for prophylaxis is still missing. However, the tegumental protein Sm29 of Schistosoma mansoni was shown to be a promising antigen to compose an anti-schistosomiasis vaccine. Though, recombinant Sm29 is expressed in Escherichia coli as insoluble inclusion bodies requiring an efficient process of refolding, thus, hampering its production in large scale. We present in this work studies to refold the recombinant Sm29 using high hydrostatic pressure, a mild condition to dissociate aggregated proteins, leading to refolding on a soluble conformation. Our studies resulted in high yield of rSm29 (73%) as a stably soluble and structured protein. The refolded antigen presented protective effect against S. mansoni development in immunized mice. We concluded that the refolding process by application of high hydrostatic pressure succeeded, and the procedure can be scaled-up, allowing industrial production of Sm29.
authors.affiliation: Centro de Biotecnologia, Instituto Pesquisas Energéticas e Nucleares, São Paulo, SP, Brazil.
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Chura-Chambi
authors.forename: Erika
authors.initials: E
authors.lastname: Nakajima
authors.forename: Roberta R
authors.initials: RR
authors.lastname: de Carvalho
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Miyasato
authors.forename: Sergio C
authors.initials: SC
authors.lastname: Oliveira
authors.forename: Ligia
authors.initials: L
authors.lastname: Morganti
authors.forename: Elizabeth A L
authors.initials: EA
authors.lastname: Martins
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology*; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D015801:Helminth Proteins / Q000276:immunology*; D006801:Humans; D002479:Inclusion Bodies / Q000235:genetics / Q000473:pathology; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012547:Schistosoma / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D012552:Schistosomiasis / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D014612:Vaccines / Q000096:biosynthesis*
chemical_list: D000947:Antigens, Helminth; D015801:Helminth Proteins; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C000589456:Sm29 protein, Schistosoma mansoni; D014612:Vaccines
keywords: CD; GSSG and GSH; GdnHCl; HHP; Hydrostatic pressure; IB; IMAC; LS; Protein refolding; Schistosomiasis; Vaccine; circular dichroism; guanidine hydrochloride; high hydrostatic pressure; immobilized metal affinity chromatography; inclusion bodies; light scattering; oxidized and reduced form of glutathione* respectively